US 12,433,803 B2
Methods and compositions for studying cell evolution
Pratiti Bandopadhayay, Boston, MA (US); Rameen Beroukhim, Boston, MA (US); Paul Blainey, Cambridge, MA (US); David Feldman, Cambridge, MA (US); Cory Johannessen, Cambridge, MA (US); and Funien Tsai, Cambridge, MA (US)
Assigned to THE BROAD INSTITUTE, INC., Cambridge, MA (US); DANA-FARBER CANCER INSTITUTE, INC., Boston, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
Filed by THE BROAD INSTITUTE, INC., Cambridge, MA (US); DANA-FARBER CANCER INSTITUTE, INC., Boston, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
Filed on Nov. 22, 2022, as Appl. No. 18/057,809.
Application 18/057,809 is a continuation of application No. 16/760,906, granted, now 11,547,614, previously published as PCT/US2018/058519, filed on Oct. 31, 2018.
Claims priority of provisional application 62/579,858, filed on Oct. 31, 2017.
Prior Publication US 2023/0083163 A1, Mar. 16, 2023
Int. Cl. A61F 13/551 (2006.01); A41D 19/00 (2006.01); C12N 15/10 (2006.01); C12N 15/113 (2010.01)
CPC A61F 13/5515 (2013.01) [A41D 19/002 (2013.01); A41D 19/0075 (2013.01); A61F 13/5518 (2013.01); C12N 15/1079 (2013.01); C12N 15/1082 (2013.01); C12N 15/113 (2013.01); A41D 19/0062 (2013.01); A41D 2400/52 (2013.01); A61F 2013/55155 (2013.01); C12N 2310/20 (2017.05); C12N 2320/10 (2013.01)] 20 Claims
 
1. An isolated subpopulation of originating cells of an originating cell population, wherein:
(A) each originating cell of the originating cell population comprises DNA construct comprising a unique barcode sequence integrated into the genome of each originating cell, wherein: (i) progeny cells of each originating cell of the originating cell population comprise the same barcode; and (ii) each barcode sequence comprises a guide sequence configured to guide a CRISPR-Cas effector protein to one or more target loci of a barcode sequence-matched reporter construct, wherein the barcode sequence is configured to introduce an insertion or deletion or a functional domain to the barcode sequence-matched reporter construct, thereby activating at least one of the one or more selection markers or reporters of the barcode sequence-matched reporter construct; and
(B) the isolated subpopulation of originating cells comprises the barcode sequence-matched reporter construct for one or more selected unique barcode sequences of the originating cell population and expresses the activated at least one of the one or more selection markers or reporters of the barcode sequence-matched reporter construct.